"
Team:Paris Saclay/Notebook/July/24
From 2014.igem.org
(Difference between revisions)
(→Transformation of competent E.coli cells) |
(→Plasmid DNA Extraction and Electrophoresis) |
||
| Line 6: | Line 6: | ||
====Plasmid DNA Extraction and Electrophoresis==== | ====Plasmid DNA Extraction and Electrophoresis==== | ||
| - | |||
''by Terry'' | ''by Terry'' | ||
| Line 12: | Line 11: | ||
Biobricks used: | Biobricks used: | ||
| - | + | # BBa_K1033927 with pSB1C3 (pink chromoprotein) | |
| - | + | # BBa_K1033910 with pSB1C3 (yellow chromoprotein) | |
| - | + | # BBa_K1033921 with pSB1C3 (red chromoprotein) | |
| - | + | # BBa_K1033922 with pSB1C3 (red chromoprotein) | |
| - | + | # BBa_K1033902 with pSB1C3 (blue chromoprotein) | |
| - | + | # BBa_K731201 with pSB1C3 (Arabinose inductible araC-pBAD promoter) | |
| - | + | # BBa_K1033925 with pSB1C3 (pink chromoprotein) | |
| - | + | # BBa_K1033905 with pSB1C3 (purple chromoprotein) | |
| - | + | # BBa_K1033913 with pSB1C3 (orange chromoprotein) | |
| - | + | # BBa_K1033929 with pSB1C3 (blue chromoprotein) | |
| + | |||
| + | '''Result:''' | ||
| + | |||
| + | Success, all DNAs have the expected size | ||
[https://2014.igem.org/Team:Paris_Saclay/Protocols/Extraction_of_the_Genomic_DNA_from_Bacteria_by_using_NucleoSpin%C2%AE_Tissue Protocol] | [https://2014.igem.org/Team:Paris_Saclay/Protocols/Extraction_of_the_Genomic_DNA_from_Bacteria_by_using_NucleoSpin%C2%AE_Tissue Protocol] | ||
Revision as of 15:02, 25 July 2014
Contents |
Thursday 24th July
Lab work
B - Construction of the fusion protein (color)
Plasmid DNA Extraction and Electrophoresis
by Terry
Biobricks used:
- BBa_K1033927 with pSB1C3 (pink chromoprotein)
- BBa_K1033910 with pSB1C3 (yellow chromoprotein)
- BBa_K1033921 with pSB1C3 (red chromoprotein)
- BBa_K1033922 with pSB1C3 (red chromoprotein)
- BBa_K1033902 with pSB1C3 (blue chromoprotein)
- BBa_K731201 with pSB1C3 (Arabinose inductible araC-pBAD promoter)
- BBa_K1033925 with pSB1C3 (pink chromoprotein)
- BBa_K1033905 with pSB1C3 (purple chromoprotein)
- BBa_K1033913 with pSB1C3 (orange chromoprotein)
- BBa_K1033929 with pSB1C3 (blue chromoprotein)
Result:
Success, all DNAs have the expected size
C - Lemon scent
Plasmid DNA Extraction & Electrophoresis
by Terry
Biobricks used:
- 7. BBa_K762100 with pSB1C3 (Limonen synthase1 with RBS)
Gel electrophoresis of p cola
by Sean
p cola used: sample left overnight yesterday resuspended in 30μl of H2O milliQ.
Results
Results of the PCR Targeting
by Romain
PCR targeting made the July 22nd. Strains used: DY330 (clone I and II). Plasmid used: pJBEI-6409 (code for enzymes to produce limonene from glucose in E. coli)
Results:
- Nothing has grown on the control dishes.
- Failure, nothing has grown on the clone I dishes.
- Failure, other bacteria have grown on the clone II dishes.
We have to make another PCR. Liquid culture of DY330 made in 30ml LB.
E - Salicylate Inducible Suppressing System
Plasmid DNA Extraction & Electrophoresis
by Terry
Biobricks used:
- 12 & 13. BBa_J45017 with pSB1C3 with pSB1AK3 (pchBA synthase (converts chorismate to salicylate))
Transformation of competent E.coli cells
by Fabio
We transformed in DH5α both BioBricks rehydrated yesterday (BBa_J61051 and BBa_K228001).
Samples:
- BBa_J61051 Concentrated
- BBa_J61051 Undiluted
- BBa_K228001 Concentrated
- BBa_K228001 Undiluted
- DH5α Control
Results on 25th July morning:
- Success, grown very well (> 200 colonies)
- Success, grown well (> 50 colonies)
- Success, grown well (> 90 colonies)
- Success, grown (15 colonies)
- Success, nothing has grown
Members present:
- Instructors and advisors: Alice and Solenne.
- Students: Arnaud, Fabio, Juliette, Pierre, Romain, Sean and Terry.
